Rapid restriction enzyme free detection of DNA methyltransferase activity based on DNA-templated silver nanoclusters

被引：0
作者：
Hanie Ahmadzade Kermani
Morteza Hosseini
Mehdi Dadmehr
Mohammad Reza Ganjali
机构：
[1] University of Tehran,Department of Life Science Engineering, Faculty of New Sciences & Technologies
[2] Payame Noor University,Department of Biotechnology
[3] University of Tehran,Center of Excellence in Electrochemistry, Faculty of Chemistry
来源：
Analytical and Bioanalytical Chemistry | 2016年 / 408卷
关键词：
DNA methyltransferase enzyme; Silver nanoclusters; Fluorescence assay; M.SssI activity;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
DNA methylation has significant roles in gene regulation. DNA methyltransferase (MTase) enzyme characterizes DNA methylation and also induces an aberrant methylation pattern that is related to many diseases, especially cancers. Thus, it is required to develop a method to detect the DNA MTase activity. In this study, we developed a new sensitive and reliable method for methyltransferase activity assay by employing DNA-templated silver nanoclusters (DNA/Ag NCs) without using restriction enzymes. The Ag NCs have been utilized for the determination of M.SssI MTase activity and its inhibition. We designed an oligonucleotide probe which contained an inserted six-cytosine loop as Ag NCs formation template. The changes in fluorescence intensity were monitored to quantify the M.SssI activity. The fluorescence spectra showed a linear decrease in the range of 0.4 to 20 U/ml with a detection limit of 0.1 U/ml, which was significant compared with previous reports. The proposed method was applied successfully for demonstrating the Gentamicin effect as MTase inhibitor. The proposed method showed convenient reproducibility and sensitivity indicating its potential for the determination of methyltransferase activity.
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页码：4311 / 4318
页数：7
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